绵羊FGF5基因的克隆、表达及RNA干扰

doi:10.3724/SP.J.1005.2011.00982

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HEREDITAS ›› 2011, Vol. 33 ›› Issue (9): 982-988.doi: 10.3724/SP.J.1005.2011.00982

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Cloning, expression analysis and RNA interference of FGF5 gene in sheep

LIU Wu-Xian¹, JIA Bin¹, SHI Guo-Qing^1,3, REN Jian-Gong², LIU Ka², MA Run-Lin²

1. College of Animal Science, Shihezi University, Shihezi 832000, China 2. Institute of Genetics and Developmental Biology, Chinese Academy of Sciences, Beijing 100101, China 3. Institute of Animal Husbandry and Veterinary Medicine, Xinjiang Academy of Agricultural and Reclamation Science, Shihezi 832000, China

Received:2010-12-03 Revised:2011-01-12 Online:2011-09-20 Published:2011-09-25

Abstract

Abstract: The cDNA of fibroblast growth factor 5 (FGF5) gene in sheep was cloned, and the nucleotides sequence homology of FGF5 was compared with other six mammal. In addition, the expression of FGF5 in different tissues was analysed. Gene FGF5 was then recombined into prokaryotic expression vector (pGEX-4T-2) and RNA interference vector (pSilencer 5.1 H1) to study its pression in fibroblast cell lines. Results showed that the open reading frame (ORF) of cDNA in sheep consisted of 813 nucleotide acids encoding 270 amino acids, with the molecular mass of 29.58 kDa and theoretical pI of 10.59. The amino acids sequence of FGF5 gene in sheep shared high identity with those in cow, human, mouse, rat, dog, cat and rabbit. In addition, analysis on tissue expression showed that FGF5 expressed in skin, heart, kidney, liver, pancreas, spleen, lung, and small intestine, especially presenting high levels in skin. The ex-pression of FGF5 in E. coli was induced with IPTG, which produced a protein band with the expected size of 56 kDa on SDS-PAGE, while the expression of FGF5 in sheep fibroblast cell line was knocked down remarkably with the help of integrated RNAi vector.

Key words: expression, RNAi, sheep, FGF5, gene cloning

LIU Wu-Wen, GU Bin, DAN Guo-Qiang, LIN Jian-Gong, LIU Ka, MA Run-Lin. Cloning, expression analysis and RNA interference of FGF5 gene in sheep[J]. HEREDITAS, 2011, 33(9): 982-988.

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Cloning, expression analysis and RNA interference of FGF5 gene in sheep

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